JP6348845B2 - Prenyloxyquinolinecarboxylic acid derivative - Google Patents

Description

  The present invention relates to a prenyloxyquinolinecarboxylic acid derivative containing a compound having an uncoupling action and / or an immunosuppressive action as an active ingredient.

  ATP production in mitochondria is performed by TCA cycle and oxidative phosphorylation reaction composed of β-oxidation system and electron transport system. A proton concentration gradient is formed in the mitochondrial inner membrane by the electron transfer system, and F-ATPase is rotated by the energy gradient to synthesize ATP (Non-patent Document 1). Thus, in mitochondrion, electron transport and ATP synthesis are closely coupled via a proton concentration gradient in the inner membrane.

  UCP (uncoupling protein), which is an uncoupling protein of the inner mitochondrial membrane, is known as a channel that eliminates a proton concentration gradient in a short-circuit manner (Non-patent Document 1). When UCP is activated, protons flow out through the inner mitochondrial membrane, which eliminates the proton concentration gradient. As a result, the chemical energy of the oxidized substrate is converted into heat energy and dissipated without being used for ATP synthesis. Therefore, if this process is repeated, consumption of intracellular fat is promoted due to a decrease in ATP production efficiency, which can be a countermeasure against obesity. Therefore, as with UCP, uncoupling agents that act on the mitochondrial inner membrane to eliminate the proton concentration gradient have attracted attention as anti-obesity agents.

  Many uncoupling agents have been discovered or synthesized so far, and their effects on cells and individuals have been published (Non-patent Documents 2 and 3). For example, dinitrophenol (DNP) has a significant weight loss effect and has been used as an anti-obesity drug, but has a serious problem that it involves serious side effects such as neuropathy and cataract (Non-Patent Document 4). In general, the effect of uncoupling agents to suppress body weight gain is useful as a measure against obesity, but normally, the uncoupling action remarkably eliminates the proton concentration gradient, thereby inhibiting ATP production and promoting active oxygen production. Therefore, there was a problem that side effects were large.

Nedergaard J., et al., 2005, EMBO reports, 6: 917-921. Terada H., 1990, Environ Health Perspect, 87: 213-218. Martinean L.C., 2012, Biochim Biophys Acta, 1820: 133-150. De Felice F.G., & Ferreira S.T., 2006, IUBMB Life, 58: 185-191.

  If an uncoupling agent that acts gently on the inner mitochondrial membrane and does not inhibit ATP synthesis is developed, it can be a weight gain inhibitor with few side effects.

  Accordingly, an object of the present invention is to develop and provide a compound having an uncoupling action with few side effects as an active ingredient of a weight gain inhibitor.

  In order to solve the above-mentioned problems, the present inventors have conducted intensive research, and as a new uncoupler, found a low molecular weight compound Ppc1 derived from Polysphondylium pseudo-candidum, which is a kind of cellular slime mold. Conventionally, Ppc1 was only known to exhibit a growth-inhibiting action on cultured cells (Kikuchi H. et al., 2010, Tetrahedron, 66: 6000-6007). The present inventors have now clarified that Ppc1 does not inhibit the mitochondrial proton concentration gradient and ATP synthesis, and that ATP production efficiency is lowered by uncoupling of protons through the inner mitochondrial membrane. Ppc1 also had the advantage of low cytotoxicity because of its mild uncoupling effect in ATP synthesis. Furthermore, a novel compound of a prenyloxyquinolinecarboxylic acid derivative having the same uncoupling action as Ppc1 has been found. Some of them have an immunosuppressive effect. The present invention has been made based on these new findings and provides the following.

［式中、R¹は、水素原子（H）又はC⁵、C¹⁰、C¹⁵若しくはC²⁰のプレニル基を表し、R²は、C¹〜C¹⁰の直鎖状若しくは分岐状のアルキル基、又はC⁵、C¹⁰、C¹⁵若しくはC²⁰のプレニル基を表し、R³及びR⁴は、それぞれ独立してC⁵、C¹⁰、C¹⁵若しくはC²⁰のプレニル基（ただし、R¹がH及びR²がメチル基のときのC⁵のプレニル基を除く）、又はイソペンチル基を表す）］(1) A compound represented by the following general formula (I) or a salt thereof.

[Wherein, R ¹ represents a hydrogen atom (H) or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, and R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group. Or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, wherein R ³ and R ⁴ are each independently a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group (where R ¹ is A C ⁵ prenyl group when H and R ² are methyl groups) or an isopentyl group)]

(2) R ¹ represents H or a dimethylallyl group, R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group, or a C ⁵ or C ¹⁰ prenyl group, R ³ and R ⁴ is independently a C ⁵ or C ¹⁰ prenyl group (except for the C ⁵ prenyl group when R ¹ is H and R ² is a methyl group), or an isopentyl group, Or a salt thereof.

(3) The compound according to (2) or a salt thereof represented by the following formulas (II) to (V):

(4) The compound according to (1) or a salt thereof represented by the following formulas (VIII) to (XII):

(5) The compound represented by the general formula (I) described in (1), wherein R ¹ represents H, or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, and R ² represents , C ¹ -C ¹⁰ linear or branched alkyl group (excluding C ² -C ⁴ alkyl group when R ¹ is H), or C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ Wherein R ³ and R ⁴ each independently represents a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group or a salt thereof as an active ingredient.

(6) R ¹ represents H or a dimethylallyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (however, a C ^{2 to} C ⁴ alkyl group when R ¹ is H) Or a prenyl group of C ⁵ or C ¹⁰ , and R ³ and R ⁴ represent a dimethylallyl group, the weight gain inhibitor according to (4).

(7) The body weight gain inhibitor according to (5), which is represented by the above formulas (II) to (V) and the following formula (VI).

(8) Use of the compound or a salt thereof according to (3) for the manufacture of a weight gain inhibitor.

(9) The compound represented by the general formula (I) described in (1), wherein R ¹ represents a hydrogen atom (H) or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group. , R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group, or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, and R ³ and R ⁴ are each independently Effective compounds such as C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl groups (excluding C ⁵ prenyl groups when R ¹ is H and R ² is a methyl group), or isopentyl groups, or salts thereof An immunosuppressant contained as a component.

(10) R ¹ represents H or a dimethylallyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (however, a C ³ or C ⁴ alkyl group when R ¹ is H) Or a compound or a salt thereof, which represents a C ⁵ or C ¹⁰ prenyl group, and R ³ and R ⁴ each independently represents a C ⁵ or C ¹⁰ prenyl group or geranyl group, or an isopentyl group. As an immunosuppressant.

(11) The immunosuppressive agent according to (9), which is represented by the above formulas (II) to (VI) and (VIII) to (XII).

(12) Use of the compound according to (1) to (4) or a salt thereof for the production of an immunosuppressant.

  This specification includes the contents described in the specification and / or drawings of Japanese Patent Application No. 2012-230985, which is the basis of the priority of the present application.

  According to the weight gain inhibitor of the present invention, it is possible to provide a weight gain inhibitor comprising as an active ingredient an uncoupling agent with few side effects on an individual.

  According to the immunosuppressive agent of the present invention, an immunosuppressive agent comprising the prenyloxyquinolinecarboxylic acid derivative of the present invention as an active ingredient can be provided.

The structural formula of the prenyloxyquinolinecarboxylic acid derivative synthesized in Example 1 and verified for the uncoupling action is shown. (A) is the compound MHQC, (b) is the compound Ppc1-r1, (c) is the compound Ppc1-r2, (d) is Ppc1, (e) is the compound Ppc1-r3, (f) is the compound Ppc1-r4, ( g) is compound XA-r1, (h) is compound XA-r2, (i) is compound XA-r3, and (j) is compound XA-r4. The uncoupling action of each prenyloxyquinolinecarboxylic acid derivative shown in FIG. 1 is shown. The oxygen consumption with time in mitochondria when Ppc1 (A) or XA-r3 (B) is added at different concentrations is shown. In the figure, the arrow indicates the point of addition of 400 μM ADP. The amount of ATP synthesized in mitochondria by the addition of Ppc1 (A) and XA-r3 (B) is shown. The amount of Cont CCCP added is 1 μM. The amount of ATP degradation in mitochondria by addition of Ppc1 (A) and XA-r3 (B) is shown. The amount of Cont CCCP added is 1 μM. It is the figure which showed the membrane potential of the mitochondria by addition of Ppc1 (A) and XA-r3 (B) by the fluorescence intensity ratio. It is the figure which showed the change of the cell growth of RPE cell by addition of Ppc1 (A) and XA-r3 (B). It is the figure which showed the cytotoxic activity of RPE cell by addition of Ppc1 (A) and XA-r3 (B) by the lactate dehydrogenase (LDH) activity. It is the figure which showed the change of the generation amount of the active oxygen in RPE cell by addition of Ppc1 (A) and XA-r3 (B). It is the figure which showed the weight change of the mouse | mouth when Ppc1 (A) and XA-r3 (B) are administered. 2 shows the amount of IL-2 produced in the presence of each prenyloxyquinolinecarboxylic acid derivative shown in FIG. 1 and the compound Ppc1-r5 represented by the formula (XII). It is a figure which shows the immunosuppressive effect of XA-r3. The titers of anti-NP-IgM antibody and anti-NP-IgG3 antibody contained in serum are shown as relative values of absorbance (OD ₄₅₀ ). Compound 55 represents XA-r3.

  The present invention will be specifically described.

1. Prenyloxyquinolinecarboxylic acid derivative A first aspect of the present invention is a derivative of 4,8-dihydroxyquinoline-2-carboxylic acid (xanthurenic acid) represented by the following general formula (I) (hereinafter referred to as “ Abbreviated to “prenyloxyquinolinecarboxylic acid derivative”) and salts thereof.

In the above formula (I), R ¹ represents a hydrogen atom (H), C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched group. Or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, wherein R ³ and R ⁴ are each independently a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group (provided that R ¹ represents H and R ² is a methyl group, except for a C ⁵ prenyl group), or an isopentyl group. The “prenyl group” in the general formula (I) is a prenyl group having 5, 10, 15 or 20 carbon atoms, specifically, a dimethylallyl group having 5 carbon atoms, a geranyl group having 10 carbon atoms, or a farnesyl group having 15 carbon atoms. Or a geranylgeranyl group having 20 carbon atoms. Among these, the prenyl group of R ¹ , R ³ and R ⁴ is preferably a dimethylallyl group, and the prenyl group of R ² is preferably a dimethylallyl group or a geranyl group.

  The “isopentyl group” is a branched functional group having 5 carbon atoms and no double bond.

  The “alkyl group” is a linear or branched alkyl group having 1 to 10 carbon atoms. Specifically, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, isopentyl, sec-pentyl Group, tert-pentyl group, neopentyl group, 2-methylbutyl group, 1,2-dimethylpropyl group, 1-ethylpropyl group, cyclopentyl group, n-hexyl group, isohexyl group, sec-hexyl group, tert-hexyl group, Neohexyl group, 2-methylpentyl group, 1,2-dimethylbutyl group, 2,3-dimethylbutyl group, 1-ethylbutyl group, cyclohexyl group, n-heptyl group, isoheptyl group, sec-heptyl group, tert-heptyl group , Neoheptyl group, cycloheptyl group, n-octyl group, isooctyl group, sec-octyl group, tert-octyl group, neooctyl group, 2-ethylhexyl group, cyclooctyl group, n-nonyl group Isononyl group, sec- nonyl group, tert- nonyl, neononyl group, cyclononyl group, n- decyl group, an isodecyl group, sec- decyl group, tert- decyl, neodecyl group, cyclodecyl group, and the like. In particular, a methyl group, ethyl group, n-pentyl group, n-hexyl group, n-heptyl group, n-octyl group, n-nonyl group which is a linear alkyl group having 1, 2 or 5 to 10 carbon atoms , N-decyl group is preferred.

Of the prenyloxyquinolinecarboxylic acid derivatives of the present invention, compounds represented by the following formulas (II) to (V) having uncoupling action and immunosuppressive action described later are particularly preferred.

In the present specification, the compound represented by the formula (II) is referred to as “XA-r3”. The method for synthesizing XA-r3 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

In the present specification, the compound represented by the formula (III) is referred to as “XA-r4”. The method for synthesizing XA-r4 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

In the present specification, the compound represented by the above formula (IV) is referred to as “XA-r2”. The method for synthesizing XA-r2 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

  In the present specification, the compound represented by the formula (V) is referred to as “Ppc1-r4”. The method for synthesizing Ppc1-r4 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

Furthermore, among the prenyloxyquinolinecarboxylic acid derivatives of the present invention, compounds represented by the following formulas (VIII) to (XII) having an immunosuppressive action described later are also preferable.

In the present specification, the compound represented by the formula (VIII) is referred to as “Ppc1-r1”. The method for synthesizing Ppc1-r1 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

In the present specification, the compound represented by the formula (IX) is referred to as “Ppc1-r2”. The method for synthesizing Ppc1-r2 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

In the present specification, the compound represented by the formula (X) is referred to as “Ppc1-r3”. The method for synthesizing Ppc1-r3 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

In the present specification, the compound represented by the formula (XI) is referred to as “XA-r1”. The method for synthesizing XA-r1 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

  In the present specification, the compound represented by the formula (XII) is referred to as “Ppc1-r5”. The method for synthesizing Ppc1-r5 is not particularly limited. For example, it can be chemically synthesized using the synthesis methods described in the examples below.

  The “salt of prenyloxyquinolinecarboxylic acid derivative” in the present invention refers to a salt of a compound represented by the formula (I) and an acid addition salt of an active compound prepared using an acid. A pharmaceutically acceptable non-toxic acid addition salt is preferred.

  Examples of acid addition salts include inorganic acid salts such as hydrochloride, sulfate, nitrate, phosphate, carbonate, bicarbonate or perchlorate, acetate, propionate, lactate, and maleic acid. Organic acid salts such as salts, fumarate, tartrate, malate, citrate or ascorbate, such as methanesulfonate, isethionate, benzenesulfonate or p-toluenesulfonate Examples include sulfonic acid salts or acidic amino acids such as aspartate and glutamate.

  The prenyloxyquinolinecarboxylic acid derivative of the present invention has an IL-2 production inhibitory action. Therefore, it can function as an immunosuppressant.

  Also, some prenyloxyquinolinecarboxylic acid derivatives of the present invention show little cytotoxic effect and do not inhibit the mitochondrial proton concentration gradient. Furthermore, ATP production efficiency can be lowered by allowing protons to pass through the inner mitochondrial membrane by uncoupling action, and the amount of active oxygen generated at that time is very small. Therefore, it can function as an uncoupling agent with a mild uncoupling action and few side effects on the living body.

2. Weight gain inhibitor The second aspect of the present invention is a weight gain inhibitor. The weight gain inhibitor of the present invention comprises the prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof having an uncoupling action.

The “prenyloxyquinolinecarboxylic acid derivative of the first embodiment having an uncoupling action” refers to a compound represented by the above general formula (I), wherein R ¹ is H, or C ⁵ , C ¹⁰ , C ¹⁵ Or a C ²⁰ prenyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (excluding a C ^{2 to} C ⁴ alkyl group when R ¹ is H), Or it represents a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, and R ³ and R ⁴ are each independently a compound representing a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group. Preferably, R ¹ represents H or a dimethylallyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (however, a C ^{2 to} C ⁴ alkyl group when R ¹ is H) Or a prenyl group of C ⁵ or C ¹⁰ and R ³ and R ⁴ are dimethylallyl groups. Specifically, it is a compound represented by the above formulas (II) to (V) and a compound represented by the following formula (VI).

  The compound represented by the above formula (VI) is the aforementioned known compound “Ppc1”. The method for synthesizing Ppc1 is not particularly limited. An example is the method of Kikuchi et al. (Kikuchi H. et al., 2010, Tetrahedron, 66: 6000-6007).

  The prenyloxyquinolinecarboxylic acid derivative or salt thereof according to the first aspect having an uncoupling action acts as an active ingredient of the weight gain inhibitor of the present invention. The weight gain inhibitor of the present invention contains at least one prenyloxyquinolinecarboxylic acid derivative or a salt thereof according to the first aspect having an uncoupling action. Here, the salt of the prenyloxyquinolinecarboxylic acid derivative may be in the form of a prodrug in the weight gain inhibitor. As used herein, a “prodrug” is a compound that readily undergoes a chemical change under physiological conditions, resulting in a change to the prenyloxyquinolinecarboxylic acid derivative of the first aspect or an active form thereof. For example, before administration, the compound is different from the compound represented by the above general formula (I), but is converted into the compound represented by the above general formula (I) or its active form by the action of digestive enzymes in the digestive tract. Says what is done. Alternatively, it may include those that are converted to the compounds by chemical or biochemical methods in an ex vivo environment.

  The content of the prenyloxyquinolinecarboxylic acid derivative or salt thereof in the weight gain inhibitor of the present invention is the kind of prenyloxyquinolinecarboxylic acid derivative and / or its effective amount, dosage form, type of carrier to be added and administration. It depends on the form, and is appropriately selected for each condition. Usually, it is preferable that an effective amount of a prenyloxyquinolinecarboxylic acid derivative or a salt thereof is contained in one dosage unit.

  In the present specification, the “effective amount” means an amount necessary for the active ingredient to exhibit its function, that is, in the present invention, the prenyloxyquinolinecarboxylic acid derivative or a salt thereof has a weight gain inhibitory activity and / or described later. It is an amount necessary for exerting the immunosuppressive activity described in the third aspect, and refers to an amount that imparts little or no harmful side effects to the administered individual. The effective amount of the prenyloxyquinolinecarboxylic acid derivative or salt thereof is specifically 0.01 mg / kg BW (body weight) to 0.15 mg / kg BW, preferably 0.02 mg / kg BW to 0.1 mg / kg BW. As an example, for example, in the case of a tablet administered to an adult with a body weight of 60 kg a day, it may contain 0.6 mg to 9 mg of a prenyloxyquinolinecarboxylic acid derivative or a salt thereof per tablet.

  The weight gain inhibitor of the present invention can include a pharmaceutically acceptable carrier and / or solvent in addition to the prenyloxyquinolinecarboxylic acid derivative or salt thereof as an active ingredient.

  As used herein, “pharmaceutically acceptable carrier” refers to non-toxic excipients, binders, disintegrants, fillers, emulsifiers, flow additive regulators, and the like that can be commonly used in the field of pharmaceutical technology.

  Excipients include, for example, sugars (including but not limited to glucose, sucrose, lactose, raffinose, mannitol, sorbitol, inositol, dextrin, maltodextrin, starch and cellulose), metal salts (eg, sodium chloride , Sodium phosphate, calcium phosphate, calcium sulfate, magnesium sulfate, calcium carbonate), citric acid, tartaric acid, glycine, low, medium, high molecular weight polyethylene glycol (PEG), pluronic, kaolin, silicic acid, or combinations thereof It is done.

  Examples of the binder include starch paste, syrup, glucose solution, gelatin, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, shellac and / or polyvinylpyrrolidone.

  Examples of the disintegrant include starch, lactose, carboxymethyl starch, cross-linked polyvinyl pyrrolidone, agar, laminaran powder, sodium bicarbonate, calcium carbonate, alginic acid or sodium alginate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, stearic acid. Monoglycerides or their salts are mentioned.

  Examples of the filler include the sugar and / or calcium phosphate (for example, tricalcium phosphate or calcium hydrogen phosphate).

  Examples of the emulsifier include sorbitan fatty acid ester, glycerin fatty acid ester, sucrose fatty acid ester, and propylene glycol fatty acid ester.

  Examples of the flow addition modifier and the lubricant include silicate, talc, stearate or polyethylene glycol.

  In addition to the above, the pharmaceutically acceptable carrier of the present invention may contain, as necessary, isotonic agents, lubricants, flavoring agents, solubilizers, suspending agents, diluents, surfactants, and stabilizers. Agents, absorption enhancers (eg, quaternary ammonium salts, sodium lauryl sulfate), extenders, pH adjusters, humectants (eg, glycerin, starch), adsorbents (eg, starch, lactose, kaolin, bentonite, colloids) Silica), disintegration inhibitors (eg, sucrose, stearin, cocoa butter, hydrogenated oil), coating agents, colorants, preservatives, antioxidants, flavors, flavors, sweeteners, buffers, soothing agents. Etc. can also be included.

  “Pharmaceutically acceptable solvent” is a non-toxic solvent that can be usually used in the field of pharmaceutical technology, for example, water, ethanol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, And oxyethylene sorbitan fatty acid esters. These are preferably adjusted to be isotonic with blood as necessary.

  The carrier and the solvent are mainly used for facilitating the formulation and administration, and for maintaining the dosage form and the drug effect, and may be appropriately used as necessary.

  The weight gain inhibitor of the present invention has the same and / or different pharmacology as long as the prenyloxyquinolinecarboxylic acid derivative or salt thereof as an active ingredient does not lose the activity as an uncoupler, that is, the pharmacological effect of the weight gain inhibitor. One or more drugs having an effect can also be contained. For example, if the weight gain inhibitor of the present invention is an oral administration agent, a predetermined amount of a gastric mucosa protective agent can be contained as necessary.

  The weight gain inhibitor of the present invention can be formulated by a method known in the art using the prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof. For example, the method described in Remington's Pharmaceutical Sciences (Merck Publishing Co., Easton, Pa.) May be used.

  The dosage form of the weight gain inhibitor is appropriately selected depending on the administration method and / or prescription conditions. The administration method can be roughly classified into oral administration and parenteral administration.

  Examples of dosage forms suitable for oral administration include tablets, pills, granules, powders, capsules, drops, sublinguals, lozenges, liquids and the like. If necessary, the tablet can be a tablet coated with a coating known in the art, for example, a sugar-coated tablet, a gelatin-encapsulated tablet, an enteric tablet, a film-coated tablet, a double tablet or a multilayer tablet.

  In the case of oral administration, the shape and size of each dosage form are not particularly limited as long as they are within the range known in the art.

  Suitable dosage forms for parenteral agents include, for example, liquids (including suspensions), emulsions (creams), gels, ointments (including pastes), plasters, powders, or suppositories. Can be mentioned. These can be made into dosage forms suitable for the method of administration, such as systemic administration, local administration or rectal administration.

  Examples of dosage forms suitable for systemic administration include solutions as injections. Examples of dosage forms suitable for topical administration include solutions as eye drops or nasal drops, emulsions, powders as nasal drops, pastes, gels, ointments, plasters and the like. . Suitable dosage forms for rectal administration include, for example, suppositories.

  The preferred administration method in the present invention is systemic administration by injection in oral administration or parenteral administration. In the case of injection, the injection site is not particularly limited, but is injection into a blood vessel such as intravenously or intraarterially. This is because the active ingredient can be distributed throughout the body through the bloodstream, and the invasiveness to the subject is relatively low.

  The weight gain inhibitor of the present invention can suppress the weight gain of a living body regardless of whether it is an obese individual or a normal weight individual, and can bring about a weight loss effect. Therefore, the weight gain inhibitor of the present invention has an effect as an anti-obesity agent.

3. Immunosuppressive agent The third aspect of the present invention is an immunosuppressive agent. The immunosuppressive agent of the present invention comprises the prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof having an immunosuppressive action. Preferably, in the general formula (I), R ¹ represents H or a dimethylallyl group, and R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (provided that when R ¹ is H) C ³ -C ⁴ alkyl group), or a C ⁵ or C ¹⁰ prenyl group, and R ³ and R ⁴ contain a dimethylallyl group, a geranyl group, or an isopentyl group, or a salt thereof. Features. Specifically, for example, compounds represented by the above formulas (II) to (VI) and (VIII) to (XII).

  The prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof acts as an active ingredient of the immunosuppressive agent of the present invention. The immunosuppressive agent of the present invention contains at least one prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof. Here, the salt of the prenyloxyquinolinecarboxylic acid derivative may be in the form of a prodrug in the weight gain inhibitor.

  The content of the prenyloxyquinolinecarboxylic acid derivative or salt thereof in the immunosuppressive agent of the present invention is the kind of prenyloxyquinolinecarboxylic acid derivative and / or its effective amount, dosage form, type of carrier to be added, and administration form. Depending on the conditions, the conditions are appropriately selected. Usually, it is preferable that an effective amount of a prenyloxyquinolinecarboxylic acid derivative or a salt thereof is contained in one dosage unit.

  The immunosuppressive agent of the present invention can include a pharmaceutically acceptable carrier and / or solvent in addition to the active ingredient prenyloxyquinolinecarboxylic acid derivative or a salt thereof.

  The immunosuppressive agent of the present invention contains one or more drugs having the same and / or different pharmacological effects as long as the active ingredient prenyloxyquinolinecarboxylic acid derivative or salt thereof does not lose its pharmacological effect as an immunosuppressive agent. You can also For example, when the immunosuppressive agent of the present invention is an oral administration agent, a predetermined amount of a gastric mucosa protective agent can be contained as necessary.

  The immunosuppressive agent of the present invention can be formulated by a method known in the art using the prenyloxyquinolinecarboxylic acid derivative of the first aspect or a salt thereof. For example, the dosage form and administration method may be in accordance with the dosage form and administration method of the weight gain inhibitor described in the second aspect.

<Example 1: Synthesis of prenyloxyquinolinecarboxylic acid derivative>
Ten prenyloxyquinolinecarboxylic acid derivatives shown in FIG. 1 used in the examples were chemically synthesized.

(1) Synthesis of 4-methoxy-8-hydroxyquinoline-2-carboxylic acid (abbreviated herein as “MHQC”) (FIG. 1 (a): Formula VII)

  200 mg of xanthurenic acid was suspended in 2 mL of thionyl chloride and stirred at 100 ° C. After 10 hours, the temperature was returned to room temperature, and the solvent was distilled off under reduced pressure. The residue was dissolved in 2 mL of methanol and stirred at 100 ° C. After 16 hours, the temperature was returned to room temperature, and the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography, and 79 mg of methyl 4-methoxy-8-hydroxyquinoline-2-carboxylate was obtained from the fraction eluted with hexane-ethyl acetate (3: 1).

  73 mg of methyl 4-methoxy-8-hydroxyquinoline-2-carboxylate was dissolved in 7 mL of methanol, 0.7 mL of 3 M aqueous sodium hydroxide solution was added, and the mixture was stirred at 80 ° C. After 3 hours, the temperature was returned to room temperature, the reaction solution was poured into 5 mL of 1 M HCl, and extracted three times with 10 mL of chloroform. All the extracted chloroform layers were combined, washed with 10 mL of water, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography, and 66 mg of MHQC was obtained from the fraction eluted with chloroform-methanol (2: 1).

  The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. MHQC EIMS and NMR results were consistent with those described in the literature (du Moulinet D'Hardemare, A. et al., 2004, BioMetals 17: 691-697).

(2) Synthesis of Ppc1 (FIG. 1 (d): Formula VI)
62 mg of MHQC prepared in (1) was dissolved in 6 mL of N, N-dimethylformamide, 225 mg of K ₂ CO ₃ was added, and the mixture was stirred at room temperature. After 20 minutes, 0.120 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 12 hours, the reaction solution was poured into 30 mL of 0.5 M HCl and extracted three times with 30 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 30 mL of water and 30 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 65 mg of Ppc1 was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

  The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The EIMS and NMR results of Ppc1 were consistent with those described in the literature (Kikuchi H. et al., 2010, Tetrahedron, 66: 6000-6007).

(3) Synthesis of Ppc1-r1 (FIG. 1 (b): Formula VIII)
Dissolve 7 mg 3-methylbut-2-enyl 8-hydroxy-4-methoxyquinoline-2-carboxylate in 1 mL N, N-dimethylformamide, add 34 mg K ₂ CO ₃ and stir at room temperature . After 15 minutes, 0.020 mL of 1-bromo-3-methylbutane was added and stirred at room temperature. After 30 minutes, the mixture was heated to 60 ° C. and further stirring was continued. After 1 hour, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 6 mg of Ppc1-r1 was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ ・ 7.55 (1H, dd, J = 8.5, 1.2 Hz), ・ 7.60 (1H, s), 7.49 (1H, dd, J = 8.5, 7.9 Hz), 7.08 (1H, dd, J = 7.9, 1.2 Hz), 5.55-5.61 (1H, m), 4.94 (2H, d, J = 7.8 Hz), 4.22 (2H, t, J = 6.8 Hz), 4.11 (3H, s), 1.92-2.01 (3H, m), 1.80 (3H, s), 1.79 (3H, s), 1.04 (6H, d, J = 7.0 Hz).
EIMS ・ m / z (rel.int) ・ 357 [M] ⁺ (1), 300 (26), 288 (6), 246 (21), 232 (100), 219 (14).

(4) Chemical synthesis of Ppc1-r2 (Fig. 1 (c) Formula IX)
10 mg of MHQC was dissolved in 1 mL of N, N-dimethylformamide, 40 mg of NaHCO ₃ was added, and the mixture was stirred at room temperature. After 15 minutes, 0.040 mL of 1-bromo-3-methylbutane was added and stirred at room temperature. After 6 hours, the mixture was heated to 60 ° C. and further stirring was continued. After 3 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was dissolved in 1 mL of N, N-dimethylformamide, 23 mg of K ₂ CO ₃ was added, and the mixture was stirred at room temperature. After 15 minutes, 0.020 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 3 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 3 mg of Ppc1-r2 was obtained from the fraction eluted with hexane-ethyl acetate (19: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ ・ 7.76 (1H, dd, J = 8.2, 0.9 Hz), ・ 7.59 (1H, s), 7.49 (1H, t, J = 8.2 Hz), 7.08 (1H , dd, J = 8.2, 0.9 Hz), 5.67-5.73 (1H, m), 4.80 (2H, d, J = 6.9 Hz), 4.46 (2H, t, J = 7.0 Hz), 4.11 (3H, s) , 1.81 (3H, s), 1.78 (3H, s), 1.76-1.90 (3H, m), 1.00 (6H, d, J = 7.2 Hz).
EIMS ・ m / z (rel.int) ・ 357 [M] ⁺ (10), 328 (4), 289 (100), 219 (61), 201 (17), 173 (24).

(5) Synthesis of Ppc1-r3 (FIG. 1 (e): Formula X)
40 mg of MHQC was dissolved in 1 mL of N, N-dimethylformamide, 153 mg of NaHCO ₃ was added, and the mixture was stirred at room temperature. After 30 minutes, 0.040 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 15 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 33 mg of 3-methylbut-2-enyl 8-hydroxy-4-methoxyquinoline-2-carboxylate was obtained from the fraction eluted with chloroform-methanol (19: 1).

Dissolve 8 mg 3-methylbut-2-enyl 8-hydroxy-4-methoxyquinoline-2-carboxylate in 1 mL N, N-dimethylformamide, add 36 mg K ₂ CO ₃ and stir at room temperature . After 30 minutes, 0.020 mL of geranyl bromide was added and stirred at room temperature. After 90 minutes, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 6 mg of Ppc1-r3 was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ ・ 7.76 (1H, dd, J = 8.1, 1.1 Hz), ・ 7.60 (1H, s), 7.48 (1H, t, J = 8.1 Hz), 7.08 (1H , dd, J = 8.1, 1.1 Hz), 5.67-5.73 (1H, m), 5.55-5.61 (1H, m), 5.08-5.14 (1H, m), 4.95 (2H, d, J = 7.7 Hz), 4.85 (2H, d, J = 6.3 Hz), 4.11 (3H, s), 2.04-2.20 (4H, m), 1.80 (3H, s), 1.78 (6H, s), 1.65 (3H, s), 1.61 (3H, s).
EIMS ・ m / z (rel.int) ・ 423 [M] ⁺ (1), 354 (29), 287 (21), 219 (100).

(6) Chemical synthesis of Ppc1-r4 (FIG. 1 (f): Formula V)
62 mg of MHQC was dissolved in 6 mL of N, N-dimethylformamide, 225 mg of K ₂ CO ₃ was added, and the mixture was stirred at room temperature. After 20 minutes, 0.120 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 12 hours, the reaction solution was poured into 30 mL of 0.5 M HCl and extracted three times with 30 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 30 mL of water and 30 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 8 mg of Ppc1-r4 was obtained from the fraction eluted with hexane-ethyl acetate (19: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR of Ppc1-r4 are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ ・ ・ 7.59 (1H, s), 7.27 (1H, d, J = 8.1 Hz), 6.98 (1H, d, J = 8.1 Hz), 5.67-5.73 (1H , m), 5.55-5.61 (1H, m), 5.31-5.35 (1H, m), 4.94 (2H, d, J = 7.3 Hz), 4.76 (2H, d, J = 6.9 Hz), 4.05 (3H, s), 3.89 (2H, d, J = 7.2 Hz), 1.80 (6H, s), 1.78 (3H, s), 1.77 (3H, s), 1.75 (3H, s), 1.74 (3H, s).
EIMS ・ m / z (rel.int) ・ 423 [M] ⁺ (3), 355 (26), 287 (100), 219 (62).

(7) Chemical synthesis of XA-r1 (FIG. 1 (g): Formula XI)
50 mg of xanthurenic acid was suspended in 1 mL of thionyl chloride and stirred at 100 ° C. After 10 hours, the temperature was returned to room temperature, and the solvent was distilled off under reduced pressure. The residue was dissolved in 2 mL of ethanol and stirred at 100 ° C. After 24 hours, the temperature was returned to room temperature, and the solvent was distilled off under reduced pressure. The residue was subjected to silica gel column chromatography, and 27 mg of ethyl 4-ethoxy-8-hydroxyquinoline-2-carboxylate was obtained from the fraction eluted with hexane-ethyl acetate (4: 1).

27 mg of ethyl 4-ethoxy-8-hydroxyquinoline-2-carboxylate was dissolved in 2.5 mL of ethanol, 0.3 mL of 3 M aqueous sodium hydroxide solution was added, and the mixture was stirred at 80 ° C. After 4 hours, the temperature was returned to room temperature, the reaction solution was poured into 5 mL of 1 M HCl, and extracted three times with 10 mL of chloroform. All the extracted chloroform layers were combined, washed with 10 mL of water, dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The residue was dissolved in 1 mL of N, N-dimethylformamide, 135 mg of K ₂ CO ₃ was added, and the mixture was stirred at room temperature. After 20 minutes, 0.040 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 18 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 5 mg of XA-r1 was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, acetone-d ₆ ) ・ ・ 7.79 (1H, dd, J = 8.2, 1.1 Hz), ・ 7.55 (1H, s), 7.54 (1H, dd, J = 8.2, 7.9 Hz) , 7.25 (1H, dd, J = 7.9, 1.1 Hz), 5.67-5.73 (1H, m), 5.48-5.54 (1H, m), 4.90 (2H, d, J = 7.2 Hz), 4.86 (2H, d , J = 7.0 Hz), 4.41 (2H, q, J = 7.1 Hz), 1.82 (3H, s), 1.80 (3H, s), 1.79 (6H, s), 1.57 (3H, t, J = 7.1 Hz ).
EIMS ・ m / z (rel.int) ・ 369 [M] ⁺ (2), 300 (28), 272 (2), 233 (100), 205 (7).

(8) Chemical synthesis of XA-r3 (FIG. 1 (i): Formula II)
61 mg of xanthurenic acid (Aldrich, Cat. No. D120804) was dissolved in 6 mL of N, N-dimethylformamide, 400 mg of K ₂ CO ₃ was added, and the mixture was stirred at room temperature. After 30 minutes, 0.137 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 30 minutes, the mixture was heated to 40 ° C. and further stirring was continued. After 12 hours, the reaction solution was poured into 30 mL of 1M HCl and extracted three times with 30 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 30 mL of water and 30 mL of saturated brine, dried over anhydrous magnesium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 50 mg of XA-r3 was obtained from the fraction eluted with hexane-ethyl acetate (4: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) 7.79 (1H, d, J = 8.2 Hz), 7.60 (1H, s), 7.46 (1H, t, J = 8.2 Hz), 7.07 (1H, d , J = 8.2 Hz), 5.68-5.74 (1H, m), 5.55-5.63 (2H, m), 4.95 (2H, d, J = 7.2 Hz), 4.77-4.83 (4H, m), 1.84 (3H, s), 1.81 (3H, s), 1.80 (6H, s), 1.78 (6H, s).
EIMS ・ m / z (rel.int) ・ 409 [M] ⁺ (2), 340 (11), 272 (28), 205 (100).

(9) Chemical synthesis of XA-r2 (FIG. 1 (h): Formula IV)
100 mg of xanthurenic acid was dissolved in 10 mL of N, N-dimethylformamide, 508 mg of K ₂ CO ₃ was added, and the mixture was stirred at 0 ° C. After 20 minutes, 0.226 mL of 1-bromo-3-methyl-2-butene was added and stirred at 0 ° C. After 4 hours, the reaction solution was poured into 60 mL of 0.5 M HCl and extracted three times with 60 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 60 mL of water and 60 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 19 mg 3-methylbut-2-enyl 4-hydroxy-8- (3-methylbut-2-enyloxy) quinoline from the fraction eluted with hexane-ethyl acetate (3: 2) -2-carboxylate was obtained.

Dissolve 9 mg 3-methylbut-2-enyl 4-hydroxy-8- (3-methylbut-2-enyloxy) quinoline-2-carboxylate in 1 mL N, N-dimethylformamide, 35 mg K ₂ CO ₃ was added and stirred at room temperature. After 30 minutes, 0.020 mL of 1-bromopentane was added and stirred at room temperature. After 30 hours, the mixture was heated to 60 ° C. and further stirring was continued. After 2 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 7 mg of XA-r2 was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ 7.88 (1H, d, J = 7.9 Hz), ・ 7.58 (1H, s), 7.48 (1H, t, J = 7.9 Hz), 7.08 (1H, d , J = 7.9 Hz), 5.67-5.73 (1H, m), 5.53-5.60 (1H, m), 4.95 (2H, d, J = 7.6 Hz), 4.81 (2H, d, J = 7.0 Hz), 4.26 (2H, t, J = 6.9 Hz), 1.96 (2H, quint, J = 6.9 Hz), 1.80 (3H, s), 1.79 (3H, s), 1.75 (6H, s), 1.50-1.59 (2H, m), 1.41-1.50 (2H, m), 0.97 (3H, t, J = 7.1 Hz).
EIMS ・ m / z (rel.int) ・ 411 [M] ⁺ (3), 342 (36), 275 (100), 205 (37).

(10) Chemical synthesis of XA-r4 (Figure 1 (J): Formula III)
Dissolve 8 mg 3-methylbut-2-enyl 4-hydroxy-8- (3-methylbut-2-enyloxy) quinoline-2-carboxylate in 1 mL N, N-dimethylformamide, 31 mg K ₂ CO ₃ was added and stirred at room temperature. After 20 minutes, 0.020 mL of geranyl bromide was added and stirred at room temperature. After 2 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 7 mg of XA-r4 was obtained from the fraction eluted with hexane-ethyl acetate (17: 3).

The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.
¹ H-NMR (400 MHz, CDCl ₃ ) ・ ・ 7.79 (1H, d, J = 8.3 Hz), ・ 7.59 (1H, s), 7.46 (1H, t, J = 8.3 Hz), 7.07 (1H, d , J = 8.3 Hz), 5.68-5.74 (1H, m), 5.54-5.64 (2H, m), 5.08-5.14 (1H, m), 4.95 (2H, d, J = 7.4 Hz), 4.83 (2H, d, J = 6.8 Hz), 4.80 (2H, d, J = 6.7 Hz), 2.09-2.21 (4H, m), 1.81 (3H, s), 1.80 (3H, s), 1.78 (6H, s), 1.71 (3H, s), 1.68 (3H, s), 1.61 (3H, s).
EIMS ・ m / z (rel.int) ・ 477 [M] ⁺ (1), 408 (3), 341 (7), 273 (38), 205 (100).

(11) Chemical synthesis of Ppc1-r5 (Formula XII)
40 mg of MHQC was dissolved in 1 mL of N, N-dimethylformamide, 162 mg of NaHCO ₃ was added, and the mixture was stirred at room temperature. After 30 minutes, 0.040 mL of geranyl bromide was added and stirred at room temperature. After 13 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 29 mg of (E) -3,7-dimethylocta-2,6-dienyl 8-hydroxy-4- methoxyquinoline-2 was eluted from the fraction eluted with chloroform-methanol (19: 1). I got -carboxylate.

Dissolve 15 mg (E) -3,7-dimethylocta-2,6-dienyl 8-hydroxy -4-methoxyquinoline -2-carboxylate in 1 mL N, N-dimethylformamide, 70 mg K ₂ CO ₃ And stirred at room temperature. After 30 minutes, 0.020 mL of 1-bromo-3-methyl-2-butene was added and stirred at room temperature. After 3 hours, the reaction solution was poured into 5 mL of 0.5 M HCl and extracted three times with 5 mL of ethyl acetate. All the extracted ethyl acetate layers were combined, washed with 10 mL of water and 10 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography, and 8 mg (XII) was obtained from the fraction eluted with hexane-ethyl acetate (9: 1).

  The product was analyzed by mass spectrometry and NMR using the EIMS (electron impact mass spectrum) method. The results of EIMS and NMR are shown below.

¹ H-NMR (400 MHz, CDCl ₃ ) d7.75 (1H, dd, J = 7.8, 1.3 Hz), 7.60 (1H, s), 7.48 (1H, t, J = 7.8 Hz), 7.08 (1H, dd, J = 7.8, 1.3 Hz), 5.67-5.73 (1H, m), 5.55-5.61 (1H, m), 5.07-5.12 (1H, m), 4.97 (2H, d, J = 7.1 Hz), 4.80 (2H, d, J = 6.8 Hz), 4.11 (3H, s), 2.05-2.16 (4H, m), 1.80 (6H, s), 1.78 (3H, s), 1.67 (3H, s), 1.60 ( 3H, s).
EIMS m / z (rel.int) 423 [M] ⁺ (2), 355 (10), 286 (38), 219 (100).

<Example 2: Uncoupling action of prenyloxyquinolinecarboxylic acid derivative>
The uncoupling action of each prenyloxyquinolinecarboxylic acid derivative synthesized in Example 1 was verified.

(Method)
The measurement of uncoupling action is one of the respiratory states of mitochondria. In the state 4 (state 4) where the respiratory substrate adenosine diphosphate (ADP) does not exist, oxygen when prenyloxyquinolinecarboxylic acid derivative is added It was measured as an increase in consumption.

(1) Purification of mitochondrial fraction After excising the liver of a mouse (ICR, female, body weight 30 g, Nippon Claire Co., Ltd.) and perfusing with a sucrose buffer (0.25 M sucrose, 10 mM Tris-HCl pH 7.4), Nine times (v / w) sucrose buffer was added to homogenize, and centrifuged at 80 G for 10 minutes. After collecting the centrifugation supernatant, it was centrifuged again at 700 G for 5 minutes. Further, the supernatant was collected and centrifuged again at 5,000 G for 5 minutes. The precipitate obtained after centrifugation was collected and washed twice with sucrose buffer to obtain a mitochondrial fraction.

(2) Oxygen consumption measurement The mitochondrial fraction was analyzed with 100 μM palmitoyl-L-carnitine hydrochloride (Sigma-Aldrich; P1645) and fatty acid-free bovine serum albumin (BSA) so that the mitochondrial protein was 0.9 mg / mL. ) Containing 0.3% (w / v) oxygen consumption buffer (225 mM mannitol, 75 mM sucrose, 10 mM KCl, 20 mM Tris-HCl, 0.1 mM EDTA, 3 mM KH ₂ PO ₄ , pH 7.4) (Rasmussen H. & Ogata E., 1966, Biochemistry, 5: 733-745; St-Pierre J, et al., 2002, J. Biol. Chem., 277: 44784-44790). 20 μM of each prenyloxyquinolinecarboxylic acid derivative synthesized in Example 1 was added to 100 μL of the suspension. The hydroxyquinoline derivative was dissolved in DMSO and then added so that the final concentration of DMSO was 5% (v / v). The oxygen concentration of 100 μL of the suspension was measured at 30 ° C. using an oxygen electrode MT200 Respirometer Cell (Strathkelvin Instruments Limited). In this measurement, palmitoyl-L-carnitine was used as a source of hydrogen to the mitochondrial respiratory chain via β-oxidation of fatty acids (St-Pierre J, et al., 2002, J. Biol. Chem., 277: 44784-44790).

(result)
The results are shown in FIG. As shown in this figure, Ppc1, Ppc1-r4, XA-r2, XA-r3, and XA-r4 were found to have a high effect of promoting oxygen consumption in mitochondria and a strong uncoupling effect. . That is, these prenyloxyquinolinecarboxylic acid derivatives can act as uncouplers. In the following examples, each effect was verified using Ppc1 and XA-r3 among the prenyloxyquinolinecarboxylic acid derivatives in which the uncoupling activity was observed.

<Example 3: Change with time of uncoupling action in Ppc1 and XA-r3>
The prenyloxyquinolinecarboxylic acid derivatives Ppc1 and XA-r3 whose uncoupling action was confirmed in Example 2 were examined for changes over time in the uncoupling action at the addition concentration.

(Method)
The uncoupling action was measured as an increase in mitochondrial oxygen consumption when a prenyloxyquinolinecarboxylic acid derivative (here, Ppc1 or XA-r3) was added as in Example 2.

  The basic method was performed according to Example 2. The addition concentrations of Ppc1 and XA-r3 (DMSO solution) were 0 μM, 1 μM, 5 μM, and 10 μM, and the mitochondrial oxygen consumption was added and measured for 10 minutes. ADP (MP Biomedicals, Solon; 160053) was added at a final concentration of 400 μM 5 minutes after the addition of Ppc1 or XA-r3. The final concentration of DMSO was set to 5% (v / v).

(result)
The results are shown in FIG. A is the result of Ppc1, and B is the result of XA-r3. As shown in this figure, in both Ppc1 and XA-r3, in the state 4 before the addition of ADP, an increase in oxygen consumption, that is, an increase in uncoupling activity can be observed depending on the concentration of the compound. On the other hand, in the state 3 state after the addition of ADP, the promotion of oxygen consumption can be observed equally at any compound concentration, suggesting that it hardly affects ATP synthesis in mitochondria.

<Example 4: ATP synthesis of Ppc1 and XA-r3>
The effect of ATP synthesis in mitochondria by addition of Ppc1 and XA-r3 was examined.

(Method)
The synthesis of ATP consists of an oxygen consumption buffer (225 mM mannitol, 75 mM sucrose, containing 100 μM palmitoyl-L-carnitine hydrochloride and 0.3% (w / v) BSA so that the mitochondrial protein is 0.3 mg / mL. Observation was performed using a mitochondrial fraction suspended in 10 mM KCl, 20 mM Tris-HCl, 0.1 mM EDTA, 3 mM KH ₂ PO ₄ , pH 7.4). To 100 μL of this suspension, Ppc1 or XA-r3 (DMSO solution) was added to final concentrations of 0 μM, 1 μM, 2 μM, and 5 μM, and 400 μM ADP was further added. As a control, a DMSO solution of carbonyl cyanide m-chlorophenylhydrazone (CCCP), which is an uncoupler, was added to mitochondria so that the final concentration was 1 μM. In both cases, the final concentration of DMSO was set to 5% (v / v).

  After reacting at 25 ° C for 1 minute, add 10 µL of the reaction mixture to a solution containing 10 µg of ATP synthesis inhibitor oligomycin (Wako Pure Chemical Industries) per 1 mg of mitochondrial protein, and solubilize mitochondria to synthesize ATP. Stop it. The amount of ATP in the reaction mixture was quantified at 0 minutes (before the reaction) and after 1 minute, and the difference was calculated as the amount of ATP synthesis.

  The quantification of ATP was performed using a chemiluminescence quantification reagent (“Cellular” ATP measurement reagent; Toyo Benet). The amount of chemiluminescence produced was measured with a TD-20 / 20 luminometer (Promega, Madison). The concentration was determined from a calibration curve prepared using ATP (Sigma-Aldrich).

(result)
The results are shown in FIG. A is the result of Ppc1, and B is the result of XA-r3. As shown in this figure, both Ppc1 and XA-r3 were found to have little effect on ATP synthesis in mitochondria.

<Example 5: ATP degradation of Ppc1 and XA-r3>
Contrary to Example 4, the effect of ATP degradation in mitochondria by addition of Ppc1 and XA-r3 was examined.

(Method)
Degradation of ATP is as follows: ATPase buffer containing 100 μM palmitoyl-L-carnitine hydrochloride, 0.3% (w / v) BSA, 1 mM MgCl ₂ , 1 mM KCN, and 2 mM ATP so that the mitochondrial protein is 0.6 mg / mL Observation was performed using a mitochondrial fraction suspended in (225 mM mannitol, 75 mM sucrose, 10 mM KCl, 20 mM Tris-HCl, 0.1 mM EDTA, 3 mM CH ₃ COOK, pH 7.4). Ppc1 or XA-r3 (DMSO solution) was added to 200 μL of the suspension so that the final concentrations were 0 μM, 1 μM, 2 μM, and 5 μM. 1 μM CCCP was used as a control. In both cases, the final DMSO concentration was 5% (v / v). After 10 minutes of reaction at 30 ° C., 400 μL of 6.9% perchloric acid was added to stop the decomposition of ATP (see Rigoulet M., et al., 1996, Eur. J. Biochem. 241: 280-285) The inorganic phosphorus in the supernatant after centrifugation at 10,000 G was quantified by measuring the absorbance at 883 nm by the phosphomolybdic acid method (see Allen RJ, 1940, Biochem. J. 34: 858-865). The concentration difference from the sample to which perchloric acid was added before the reaction was taken as the ATP decomposition amount.

(result)
The results are shown in FIG. A is the result of Ppc1, and B is the result of XA-r3. As shown in this figure, both Ppc1 and XA-r3 were found to have little effect on ATP degradation in mitochondria.

<Example 6: Mitochondrial membrane potential of Ppc1 and XA-r3>
The effect of addition of Ppc1 and XA-r3 on mitochondrial membrane potential was examined.

(Method)
The mitochondrial fraction was suspended in an oxygen consumption buffer containing 100 μM palmitoyl-L-carnitine hydrochloride, 0.3% (w / v) BSA, 2.5 μM safranin O so that the mitochondrial protein was 0.3 mg / mL. 100 μL of the suspension was dispensed into 96-well microplates, and Ppc1 or XA-r3 (DMSO solution) was added to a final concentration of 0 μM, 1 μM, 2 μM, or 5 μM. 1 μM CCCP was used as a control. In both cases, the DMSO solution was adjusted to a final concentration of 5% (v / v). After reaction at 25 ° C. for 20 minutes, fluorescence by excitation light of 495 nm was measured at 586 nm (Votyakova TV & Reynolds IJ, 2001, J. Neurochemistry 79: 266-277). Fluorescence intensity was measured using a Varioskan Flash (Thermo Fisher Scientific, Waltham) microplate reader. The fluorescence was measured from the top surface of the microplate.

  Membrane potential is expressed by the formula of ((B−measured value) / (B−A)} × 100 (%), where A is the value of the sample not containing Ppc1 or XA-r3, and B is the value of the sample not containing mitochondria. Calculated from

(result)
The results are shown in FIG. A is the result of Ppc1, and B is the result of XA-r3. As shown in this figure, both Ppc1 and XA-r3 were found to hardly inhibit the mitochondrial membrane potential.

<Example 7: Cytotoxicity by Ppc1 and XA-r3>
The cytotoxicity by Ppc1 and XA-r3 was verified.

(Method)
(1) Culture of retinal pigment epithelium (RPE) cells FBS (Cansera International) heat-inactivated in medium (DMEM F-12 HAM, Sigma-Aldrich) with Dulbecco's modified Eagle medium plus F12 ham as a nutrient mixture Using the culture solution added to 10% (v / v), RPE cells were cultured at 37 ° C. in the presence of water vapor saturation and 5% CO ₂ . For the measurement of cell proliferation and cytotoxicity, 6 × 10 ³ cells were seeded per well and used for the experiment after 16 hours of culture.

(2) Measurement of cell proliferation and cytotoxicity Cell proliferation and cytotoxicity were measured by lactate dehydrogenase (LDH) activity (Decker T., & Lohmann-Matthes ML, 1988, J. Immunol. Meth. 115). : 61-69). The medium cultured for 16 hours was replaced with a new medium containing 0 μM, 1 μM, 2 μM or 5 μM Ppc1 or XA-r3 (DMSO solution), and further cultured for 24 hours. The DMSO solution was adjusted to a final concentration of 0.25% (v / v). After culture, LDH activity was measured using CytoTox-ONE Homogeneous Membrane Integrity Assay (Promega). The measurement was carried out by measuring the fluorescence intensity of 590 nm by excitation light of 560 nm of resorufin reduced by reduced nicotinamide adenine dinucleotide produced by LDH using lactic acid as a substrate. Since the number of living cells is proportional to CB when the measured values when the culture solution alone, the culture solution after culture, and the cells are solubilized are A, B, and C, respectively, the value of the sample with no compound added is 100% It was shown in the ratio. Cytotoxicity was calculated by {(B−A) / (C−A)} × 100 (%), and normalized with the value of the sample to which no compound was added.

(result)
FIG. 7 shows the results of cell proliferation, and FIG. 8 shows the results of cytotoxic activity. A is the result of Ppc1, and B is the result of XA-r3. As shown in FIG. 7, although the cell proliferation ability slightly decreased as the added concentrations of Ppc1 and XA-r3 increased, a remarkable cell proliferation inhibitory effect could not be confirmed. In addition, LDH activity hardly changed even when the addition concentrations of Ppc1 and XA-r3 were increased. From this result, it was revealed that Ppc1 and XA-r3 have almost no cytotoxicity.

<Example 8: Amount of generated active oxygen>
The change in the amount of active oxygen generated in the cells by addition of Ppc1 and XA-r3 was examined.

(Method)
RPE cells were cultured according to Example 7. A 96-well microplate was seeded with 2.5 × 10 ⁴ cells / mL RPE cells per well and cultured for 16 hours. The culture medium after the culture is replaced with a new medium added so that the final concentration of Ppc1 or XA-r3 (DMSO solution) is 0 μM, 1 μM, 2 μM or 5 μM, and the final concentration of dihydroethidium (DMSO solution) is 25 μM. Incubated for an additional 30 minutes. The DMSO solution was adjusted to a final concentration of 0.25% (v / v). After culturing, the fluorescence was measured as 567 nm fluorescence by 480 nm excitation light (see Budd SL, Castilho RF, Nicholls DG, 1997, FEBS Letters 415: 21-24). The fluorescence intensity was measured using a Varioskan Flash (Thermo Fisher Scientific, Waltham) microplate reader. The fluorescence measurement was performed from the bottom of the microplate. From the measured values, the difference in fluorescence intensity from the medium containing no cells was attributed to active oxygen.

(result)
FIG. 9 shows the result. A is the result of Ppc1, and B is the result of XA-r3. As shown in this figure, it was revealed that the fluorescence intensity hardly increased even when 5 μM Ppc1 or XA-r3 was added, and there was no significant change in the amount of active oxygen generated in the cells.

  As described above, the results of Examples 4 to 8 suggest that Ppc1 and XA-r3, which are novel uncouplers, are not as harmful to cells as DNP, and therefore have fewer side effects on living bodies. Yes.

<Example 9: Change in body weight of mice by administration of Ppc1 or XA-r3>
The change in the body weight of the mice when Ppc1 or XA-r3 was administered to the mice was examined.

(Method)
The breeding conditions of the mice (ICR, female, 25 g body weight, Nippon Claire Co., Ltd., Tokyo) were room temperature 22 ° C., lighting and turning on and off the light and darkness for 12 hours, and food and water were ingested freely. 0.2 mL of phosphate buffered saline solution containing Ppc1-DMSO solution or XA-r3-DMSO solution so that the final concentration of DMSO is 0.25% (v / v), Ppc1 is 0 mg / kg body weight per animal, XA-r3 was administered intraperitoneally once a week at 0.8 mg / kg body weight and XA-r3 at 0 mg / kg body weight, 0.4 mg / kg body weight, and 0.8 mg / kg body weight per animal. In the meantime, body weight was measured regularly in units of 0.1 g. The administration control was a 0.2 mL phosphate buffered saline solution containing only DMSO so that the final concentration was 0.25% (v / v).

(result)
The results are shown in FIG. It was revealed that administration of Ppc1 or XA-r3 significantly reduced body weight compared to the administration control. This result shows that Ppc1 or XA-r3, which is a novel uncoupler, has an effect as an active ingredient of a weight gain inhibitor.

<Example 10: Effect of prenyloxyquinolinecarboxylic acid derivative on IL-2 production>
The effect of each prenyloxyquinolinecarboxylic acid derivative synthesized in Example 1 on IL-2 production was examined.

(Method)
Mouse Jurkat cells suspended at 2 × 10 ⁵ cells / mL in RPMI 1640 medium containing 10% fetal calf serum were seeded at 100 μL per well in a 96-well plate. Concanavalin A (Sigma) dissolved in RPMI 1640 medium was added to each well to a final concentration of 40 μg / mL to stimulate the cells. Furthermore, each prenyloxyquinolinecarboxylic acid derivative shown in FIG. 1 and Ppc-r5 represented by the formula (XII) were added so that the final concentration was 10 μM. As a control, dimethyl sulfoxide (DMSO) was added at a final concentration of 0.2%. After culturing at 37 ° C. under 5% CO ₂ for 2 days, the amount of IL-2 contained in the culture supernatant was measured using a Human IL-2 ELISA Kit (Thermo Fisher Scientific). About the measuring method, the attached protocol was followed.

(result)
FIG. 11 shows the relative values of the IL-2 production amount when each prenyloxyquinolinecarboxylic acid derivative is added when the IL-2 production amount in the control (DMSO addition) is 100%. Compared to the control, all the derivatives except MHQC were confirmed to suppress IL-2 production. In particular, XA-r3 showed a remarkable inhibitory effect.

  IL-2 has a function of controlling cellular immunity. Therefore, suppression of the production amount suggests the immunosuppressive effect of the prenyloxyquinolinecarboxylic acid derivative.

<Example 11: Influence of antibody production ability of mice by administration of XA-r3>
In order to further verify the immunosuppressive effect of prenyloxyquinolinecarboxylic acid derivatives, the effect on antibody production in mice administered with XA-r3 was examined.

(Method)
1 mg / kg FK506 (immunosuppressant tacrolimus; Cell Signaling Technology), 0.5 mg / kg XA-r3, and control vehicle; Glyceryl trioctanoate (Sigma) were injected into the abdominal cavity of 10-week-old wild-type mouse C57BL / 6J In each group, 8 animals were administered. The administration was continued every 3 days for 3 weeks. Six hours after the first administration, 50 μg of a suspension of chicken γ globulin (Biosearch Technologies) labeled with nitrophenol (NP) and Inject Alum Adjuvant (Thermo Fisher Scientific) was administered intraperitoneally to immunize. Thereafter, after 1 week and 3 weeks, blood was collected from the tail, and serum was separated from the collected blood, and then the produced anti-NP-IgM antibody (SouthernBiotech) and anti-NP-IgG3 antibody (SouthernBiotech) were used by ELISA. ) Was measured (Kometani, et al., J Exp Med, 2011, 208 (7), 1447-1457).

(result)
FIG. 12 shows the titer of anti-NP-IgM antibody contained in serum one week after immunization and the titer of anti-NP-IgG3 antibody contained in serum three weeks after immunization. The titer was expressed as a relative value of absorbance (OD ₄₅₀ ). The titers of anti-NP-IgM antibody and anti-NP-IgG3 antibody in the XA-r3 administration group were significantly reduced compared to those in the vehicle administration group as a control. The degree of decrease was almost the same as that of the FK506 administration group as an immunosuppressant. From these results, it was revealed that XA-r3 has a high immunosuppressive action.

  All publications, patents and patent applications cited herein are incorporated herein by reference in their entirety.

Claims (10)

The compound or its salt shown by the following general formula (I).

[Where:
R ¹ represents a hydrogen atom (H) or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group,
R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group, or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group,
R ³ represents a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group , or an isopentyl group,
R ⁴ is a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group ( excluding the dimethylallyl group when R ¹ is H, R ² is a methyl group, and R ³ is a dimethylallyl group), or isopentyl Group (except when R ³ is an isopentyl group)] R ¹ represents H or a dimethylallyl group,
R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group, or a C ⁵ or C ¹⁰ prenyl group,
R ³ is Table prenyl group, or isopentyl group C ⁵ or C ^10,
R ⁴ is a C ⁵ or C ¹⁰ prenyl group (except R ¹ is H, R ² is a methyl group, and R ³ is a dimethylallyl group), or an isopentyl group (where R ³ Table vinegar but except in the case of isopentyl group),
The compound according to claim 1 or a salt thereof. The compound or its salt of Claim 2 shown by the following formula | equation (II)-(V).

The compound or its salt of Claim 1 shown by the following formula | equation (VIII)-(XII).

A compound represented by the following general formula (I):

Where
R ¹ represents H or a dimethylallyl group ,
R ² represents a methyl group or pentyl group, or a prenyl group of C ⁵ or C ^10,
R ³ and R ⁴ represent a dimethylallyl group
A weight gain inhibitor comprising as an active ingredient a compound represented by the following formulas (II) to (V) or formula (VI) or a salt thereof:

Use of the compound represented by formulas (II) to (VI) or a salt thereof according to claim 5 for the manufacture of a weight gain inhibitor. A compound represented by the following general formula (I):

Where
R ¹ represents a hydrogen atom (H) or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group,
R ² represents a C ^{1 to} C ¹⁰ linear or branched alkyl group, or a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group,
R ³ and R ⁴ are each an immunosuppressive agent containing, as an active ingredient, a compound or a salt thereof, each independently representing a C ⁵ , C ¹⁰ , C ¹⁵ or C ²⁰ prenyl group, or isopentyl group. R ¹ represents H or a dimethylallyl group,
R ² is a C ^{1 to} C ¹⁰ linear or branched alkyl group (excluding a C ³ or C ⁴ alkyl group when R ¹ is H), or a C ⁵ or C ¹⁰ prenyl group Represent,
The immunosuppressive agent according to claim 7 , comprising a compound or a salt thereof in which R ³ and R ⁴ represent a dimethylallyl group, a geranyl group, or an isopentyl group, or a salt thereof. A compound represented by the general formula (I) according to claim 7, comprising a compound represented by the following formulas (II) to (VI) and (VIII) to (XII) or a salt thereof as an active ingredient . The immunosuppressive agent according to claim 7 .

Use of a compound represented by the general formula (I) or a salt thereof according to claim 7 for the production of an immunosuppressive agent.

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